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Notes: In this study, Anti-Human p75 polyclonal antibody was used to immunoprecipitate p75 neurotrophin receptor from HEK-293 cell lysates. The Anti-Human p75 pAb was preadsorbed with protein A sepharose and this complex was used to immunoprecipitate p75 neurotrophin receptor complexes for Western blot analysis. (3200)

Notes: The authors investigated the role of p75 in brain-derived neurotrophic factor (BDNF)-mediated shifts from excitation to inhibition in sympathetic neuron-myocyte connections in cell culture. Rat sympathetic neurons and ventricular myocytes were co-cultured and transfected with an expression vector containing either wildtype human p75 or a ligand-binding deficient p75. p75 expression was monitored by staining with Promega's Anti-Human p75 pAb. (2472)

Notes: A protein complex of the p75NTR-associated cell death executor, the p75 neurotrophin receptor, and a protein named 14-3-3 was characterized in HEK293 cells. The p75 neurotrophin receptor was detected using a 1:10,000 dilution of Promega's Anti-Human p75 pAb in a Western blot with PVDF membranes blocked with 10% nonfat dry milk. (2370)

Notes: To examine the importance of the p75 neurotrophin receptor on TrkB- and TrkC-mediated signaling, MG87 mouse fibroblast cell lines stably transfected with rat TrkB or TrkC cDNA were then stably transfected with rat p75 cDNA. To confirm the expression of p75, extracts prepared from the stable cell lines were subjected to Western blot analysis using the Anti-Human p75 pAb. (2371)

Notes: A293 cells were transfected with expression vectors for HA-tagged TrkA, TrkB or TrkC with the rat p75 receptor. The cells were lysed and immunoprecipitated with an anti-HA antibody and the resulting material was blotted and shown to react with the Anti-Human p75 pAb. p75 was not detected when the cells were transfected with the EGF receptor and p75, then immunoprecipitated with an anti-EGF Receptor antibody. This system allowed analysis of deletion mutants of both TrkB and p75. (1447)

Notes: The Anti-Human p75 pAb was used for combined immunoperoxidase and immunogold-silver labeling with PARV antibodies. The Anti-Human p75 pAb was used at a 1:2000 dilution and recognized with biotinylated donkey anti-rabbit secondary antibody. Labeling was performed on forebrain vibratome sections of 40 microns. (1244)

Notes: Recombinant Human BDNF was used in studies of BDNF neutralization. The Anti-BDNF pAb (> or = 10µg/ml) was found to inhibit 50ng/ml of BDNF from activating autophosphorylation cascades of TrkB in TrkB-expressing NIH3T3 cells. Control IgY at up to 40µg/ml had no inhibitory effect on the autophosphorylation. The Anti-BDNF pAb was added to cultures of primary rat sympathetic neurons to neutralize any BDNF/p75NTR autocrine loop. BDNF causes a decrease in cell proliferation and apoptosis as judged by the CellTiter 96® Assay and the DeadEnd™ Colorimetric Apoptosis Detection System, respectively. The DeadEnd™ System was used in a unique way. Rather than staining the apoptotic nuclei with DAB via the streptavidin-HRP conjugate, a Cy®3-conjugated streptavidin was substituted. To compare the immunolocalization of the tyrosine hydroxylase protein and the p75NTR receptor, serial sections of mouse pineal glands were probed with an anti-tyrosine hydroxylase antibody and the Anti-Human p75 pAb. A lot of detail is provided for tissue processing prior to IHC. The Anti-Human p75 pAb was also used to detect p75 in mouse and rat pineal gland extracts via Western blotting. The name of the DeadEnd™ Colorimetric Apoptosis Detection System has been changed to DeadEnd™ Colorimetric TUNEL System. (0916)

J. Comp. Neurol.407, 77-91.
p75NTR immunoreactivity in the rat dentate gyrus is mostly within presynaptic profiles but is also found in so astrocytic and postsynaptic profiles.1999

Dougherty, K.D., Milner, T.A.

Notes: The Anti-Human p75 pAb was used for immunohistochemical detection of p75NTR in 2% paraformaldehyde-fixed 40µm vibratome sections of rat brain. Tissues processed for peroxidase staining were incubated with a 1:2000 dilution of the antisera and those processed for immunogold labeling were incubated with a 1:400 dilution of the antibody. Excellent detail is provided for both procedures. (1245)

Notes: For single labeling of cultured rat olfactory ensheathing cells, the explants were grown on coverslips and fixed in methanol prior to immunostaining with the Anti-Human p75 pAb. For double labeling with a β III tubulin antibody, the cells were fixed with 4% paraformaldehyde prior to incubation with a 1:200 dilution of the Anti-Human p75 pAb. (0268)

Notes: TRAF and p75 NTR were expressed in HEK 293T cells. In one experiment, TRAF was immunoprecipitated and the complex resolved on a gel and blotted with Promega's Anti-Human p75 pAb to demonstrate the interaction between the two proteins. (0109)

Notes: To examine the regulation of neurotrophin secretion, the release of neurotrophins from PC12 cells transfected with various neurotrophin expression vectors was quantitated. To quantatite BDNF an ELISA was developed by coating the plates with the TrkB-Fc receptor body and detecting with the Anti-Human BDNF pAb and the Anti-Chicken IgY, HRP Conjugate. There was no crossreactivity with NGF, NT-3, and NT-4/5 at 1 µg/ml. The Anti-Human p75 pAb was also used in this study. (2359)

Notes: Promega's Anti-Human p75 pAb was used to determine the distribution of rat neural crest cells in the anlage of the sympathetic ganglion chain. This immunostaining was combined with TUNEL staining of the tissue. (1408)

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